2. Key capabilities:
• Select a confocal geometry optimal for
your assay and throughput needs
• Achieve excellent image quality without
sacrificing throughput via our unique optical
path technology
• Acquire statistically relevant data quickly
with an advanced scientific CMOS detector,
enabling >3 log dynamic range
• Large field of view enables whole-well imaging
• Expand your research capabilities with
transmitted light, liquid handling, and
environmental options
Deeper insight into complex biology
Higher quality images, faster throughput and more powerful analysis
The ImageXpress Micro Confocal High-Content
Imaging System provides improved quantification
for live or fixed cell assays. This versatile imaging
system features a unique confocal technology
which allows you to explore more physiologically
relevant, complex three dimensional models
including spheroids, tissues, and whole organisms
and to generate publication quality images at
high throughput for samples in slides or one to
1536‑well microplates.
Confocal technology
at the speed of widefield
imaging
• Capture an entire well of a
384‑well plate with a single
image at 4X magnification
• Capture four wells of a 1536-
well plate in a single image at
4X magnification
• Throughput of >160K wells/day
confocal, >200K wells/day
widefield
2 ImageXpress Micro Confocal
3. AgileOptix Technology at the heart of the
ImageXpress Micro Confocal System
Software-selectable configurations right for your research
The ImageXpress Micro Confocal System features AgileOptix™ Spinning Disk Technology. Our optical
options make it easy to select and configure your system to ensure the best read for your assay.
Select a spinning disk confocal geometry matched to
your assay requirements.
Spinning Disk Geometry
60 µm Pinhole
(Single Disk)
60 µm Dual
Disk with
50 µm Slit
60 µm Dual
Disk with
42 µm Pinhole
High-sensitivity detection
Fast acquisition
>3 log dynamic range*
Widefield mode for flat biology
Most confocal applications
Highest resolution imaging
High throughput applications
*Powered by our highly responsive sCMOS sensor and advanced solid state light source.
Supports subcellular assays
through whole organism
assays
• Widest selection (> 25) of objectives
• 1X to 100X magnification
• Oil objectives with up to 1.4 NA
available
• Air objectives 0.05 to 0.95 NA
3ImageXpress Micro Confocal
4. Expand your research into a new
dimension
More relevant results with 3D assay models into complex biology
Complex, three-dimensional cellular models yield
more predictive, physiologically relevant results
versus monocultures or other two-dimensional
cellular models. Explore the complexities of these
models faster and gain better results even in
samples grown in a thick extracellular matrices
using the ImageXpress Micro Confocal. The
imaging system provides flexible imaging options
to meet your specific research needs, allowing
you to easily capture images from different sample
formats, including hanging drops and in round
or flat bottom plates and monitoring cell health
kinetics under environmental control for seconds,
minutes, hours, or days.
Clearer images and
improved quantitative
screening for
• Spheroids
• Thick tissue samples
• Zebrafish and C. elegans
• Homogenous no-wash assays
Spheroids in round bottom plates. Dose–dependent effects of selected compounds. Image montage of the 384 plate. Entire spheroid is captured with one
image, example segmentation of spheroid and 4-parametric curve fits for the number of live cells in spheroids.
Cell stained for nuclei or actin
growing in a 3D gel. Projection
image of seven planes acquired
with a 40X Plan Apo objective.
Live Cells
Concentration, nM
0.1 1 10 100 1000 10000
LiveCells
1000
750
500
250
0.0
1
Benefits
• Capture an entire spheroid
in one field-of-view at 20X
magnification
• Screen biologically relevant
3D spheroids in a 96 or 384
well format
• Use confocal imaging to
accurately detect cellular
responses
• Conserve storage space by
saving only 2D reconstructions
of the z plane images
for screening cancer therapeutics
Introduction
In recent years, there has been significant
progress in development of in vitro
aggregates of tumor cells for use as
models for in vivo tissue environments.
When seeded into a well of a low-
attachment round bottom microplate,
these aggregates will form a discrete
spheroid. Spheroids are believed to mimic
tumor behavior more effectively than
regular two dimensional (2D) cell cultures
because, much like tumors, they contain
both surface-exposed and deeply buried
cells, proliferating and non-proliferating
cells, and a hypoxic center with a well-
oxygenated outer layer of cells. Such 3D
spheroid models are being successfully
used in screening environments for
identifying potential cancer therapeutics.
Some challenges to developing robust
spheroid assays:
• Locating and focusing on the spheroid in
every well so it can be imaged in a single
field-of-view
• Optimizing the compound and staining
treatment to ensure dye penetration and
avoid disturbing the spheroid placement
• Acquiring representative images
throughout the 3D structure, minimizing
out-of-focus or background signal from
above and below the imaging plane
• Rapidly analyzing the images to
yield meaningful results from which
conclusions can be drawn.
Spheroid formation
and treatment
We used the following method to form
spheroids from cancer cell lines HCT116,
DU145, and HepG2. Cells were cultured
in flasks at 37 °C and 5% CO2
before
detaching and seeding into 96 or
384-well black plates with clear bottom
U-shaped wells (Corning 4520 and
3830, respectively) at densities of 1000-
1500 cells/well in the appropriate media
supplemented with fetal bovine serum
(FBS). Within 24 hours, a single spheroid
formed in the bottom of each well and
continued growing in size until it was
used for experimentation after 2-4 days
at 37 °C and 5% CO2
. Spheroids may
be cultured longer but the increasing
size may impede stain penetration and
imaging of the center-most cells. This
application note describes assays used to
determine the effects of the anti-cancer
compounds: etoposide, paclitaxel, and
Mitomycin C. Spheroid treatment began
by adding compounds into the wells at
Benef
• Capture
in one fie
magnific
• Screen b
3D sphe
well form
• Use conf
accurate
response
• Conserv
saving o
of the z p
for screening cancer therapeutics
Introduction
In recent years, there has been significant
progress in development of in vitro
aggregates of tumor cells for use as
models for in vivo tissue environments.
When seeded into a well of a low-
attachment round bottom microplate,
these aggregates will form a discrete
spheroid. Spheroids are believed to mimic
tumor behavior more effectively than
regular two dimensional (2D) cell cultures
because, much like tumors, they contain
both surface-exposed and deeply buried
cells, proliferating and non-proliferating
cells, and a hypoxic center with a well-
oxygenated outer layer of cells. Such 3D
spheroid models are being successfully
used in screening environments for
identifying potential cancer therapeutics.
Some challenges to developing robust
spheroid assays:
• Locating and focusing on the spheroid in
every well so it can be imaged in a single
field-of-view
• Optimizing the compound and staining
treatment to ensure dye penetration and
avoid disturbing the spheroid placement
• Acquiring representative images
throughout the 3D structure, minimizing
out-of-focus or background signal from
above and below the imaging plane
• Rapidly analyzing the images to
yield meaningful results from which
conclusions can be drawn.
Spheroid formation
and treatment
We used the following method to form
spheroids from cancer cell lines HCT116,
DU145, and HepG2. Cells were cultured
in flasks at 37 °C and 5% CO2
before
detaching and seeding into 96 or
384-well black plates with clear bottom
U-shaped wells (Corning 4520 and
3830, respectively) at densities of 1000-
1500 cells/well in the appropriate media
supplemented with fetal bovine serum
(FBS). Within 24 hours, a single spheroid
formed in the bottom of each well and
continued growing in size until it was
used for experimentation after 2-4 days
at 37 °C and 5% CO2
. Spheroids may
be cultured longer but the increasing
size may impede stain penetration and
imaging of the center-most cells. This
application note describes assays used to
determine the effects of the anti-cancer
compounds: etoposide, paclitaxel, and
Mitomycin C. Spheroid treatment began
by adding compounds into the wells at
for screening cancer therapeutics
Introduction
In recent years, there has been significant
progress in development of in vitro
aggregates of tumor cells for use as
models for in vivo tissue environments.
When seeded into a well of a low-
attachment round bottom microplate,
these aggregates will form a discrete
spheroid. Spheroids are believed to mimic
tumor behavior more effectively than
regular two dimensional (2D) cell cultures
because, much like tumors, they contain
both surface-exposed and deeply buried
cells, proliferating and non-proliferating
cells, and a hypoxic center with a well-
oxygenated outer layer of cells. Such 3D
spheroid models are being successfully
used in screening environments for
identifying potential cancer therapeutics.
Some challenges to developing robust
spheroid assays:
• Locating and focusing on the spheroid in
every well so it can be imaged in a single
field-of-view
• Optimizing the compound and staining
treatment to ensure dye penetration and
avoid disturbing the spheroid placement
• Acquiring representative images
throughout the 3D structure, minimizing
out-of-focus or background signal from
above and below the imaging plane
• Rapidly analyzing the images to
yield meaningful results from which
conclusions can be drawn.
Spheroid formation
and treatment
We used the following method to form
spheroids from cancer cell lines HCT1
DU145, and HepG2. Cells were culture
in flasks at 37 °C and 5% CO2
before
detaching and seeding into 96 or
384-well black plates with clear botto
U-shaped wells (Corning 4520 and
3830, respectively) at densities of 100
1500 cells/well in the appropriate med
supplemented with fetal bovine serum
(FBS). Within 24 hours, a single sphero
formed in the bottom of each well and
continued growing in size until it was
used for experimentation after 2-4 da
at 37 °C and 5% CO2
. Spheroids may
be cultured longer but the increasing
size may impede stain penetration an
imaging of the center-most cells. This
application note describes assays use
determine the effects of the anti-canc
compounds: etoposide, paclitaxel, and
Mitomycin C. Spheroid treatment beg
by adding compounds into the wells a
widefield
confocal
4 ImageXpress Micro Confocal
5. Improve visualization and
quantitation with 3D assay models
Confocal capability improves image clarity and data quality
Compared with widefield imaging, images of thick samples
captured with confocal have reduced background
and improved sharpness, resulting in improved image
segmentation.
Enhanced imaging of
tissues and 3D matrices
• Select specific cells of interest in
3D matrices such as neurons and
stem cells
• Reject high-background
fluorescence in thick tissue
samples
• Acquire Z-stacks easily with 3D
reconstruction capability
Clearer imaging of
whole organisms
• Large field of view enables
imaging of an entire well of a
384‑well plate with a single image
• Organisms remain in focus for the
duration of the experiment
• Perform sophisticated analysis
of images and create time-lapse
videos
Rat brain section, stained for nuclei and neural outgrowths. Images taken with a 20X
Plan Fluor ELWD objective, confocal with 60 µm pinhole. Left without segmentation
and right with image segmentation with Neurite Outgrowth module.
widefield widefieldconfocal confocal
5ImageXpress Micro Confocal
6. A complete solution for screening your
most complex biological questions
Delivers seamless workflows from image acquisition to data analysis
Live cell acquisition and analysis. HeLa cells expressing Cell Cycle Chromobody undergoing normal
cell division while being imaged in confocal mode. In G1, the cells have a homogeneous fluorescence
signal. During S phase, signal accumulates in the nucleus with formation of foci. In G2, the morphology
returns to homogenous and the cell divides. White arrow indicates cell before cell division and yellow
arrow indicates daughter cells after division event.
T0 39 min 9 hr, 19 min5 hr, 19 min 14 hr, 39 min
6 ImageXpress Micro Confocal
A world of applications that
exceeds your imagination
Acquire
MetaXpress® Software powers our ImageXpress Micro Confocal system, giving you precise
control over image acquisition and analysis, all within a unified interface.
• Acquisition wizard for the entire workflow avoids image import/export steps
• Laser-based and software configurable image-based auto-focusing system ensures
robust focus across a range of sample types
• Acquisition of live cell images enables monitoring of cell growth, death, differentiation,
and migration; viral or bacterial invasion, cancer metastasis, chemotaxis, drug toxicity,
or translocation
Enjoy the benefits of a streamlined high-content screening (HCS) workflow in a fully integrated environment
with our complete imaging solution for your most complex biological questions.
7. Analyze
Avoid delays in image analysis and data processing using our
MetaXpress Software with application modules that allow you to
quickly and easily analyze your data.
• Plug-and-play application modules can be adapted to hundreds
of image-based analysis workflows
• Custom module editor empowers you to further tailor your
image analysis routines for a perfect fit
• Adaptive Background Correction™ adjusts image segmentation
to the local intensity ranges and features within and between
cells for better quantitation
• 2D projection algorithms include best focus, maximum and
minimum, and sum projection for easy interpretation of 3D
image data
• Save as cell-by-cell and/or image-by-image data
AcuityXpress™ Informatics Software, data visualization, mining and
hit selection are ready to use upon system installation.
Store
Regardless of the acquisition system used, images taken can be
stored in the secure MDCStore™ Data Management Solution.
• Accessible for sophisticated analysis by the MetaXpress® High
Content Image Acquisition and Analysis Software
• Data migration portal for integration with third-party imaging
systems or analysis tools to external host databases or
third‑party applications
7ImageXpress Micro Confocal